Salicyclic acid o-hydroxyphenylamides for combating bacteria and fungi

ABSTRACT

THIS INVENTION PROVIDES A METHOD OF USE FOR COMBATING HARMFUL MICROORGANISMS, SUCH AS FUNGI AND BACTERIA, BY APPLYING A SALICYLIC O-HYDROXYPHENYLAMIDE OF THE GENERAL FORMULA   R-CO-NH-R&#39;&#39;   WHEREIN R AND R&#39;&#39; EACH REPRESENT A PHENYL GROUP WITH AN ORTHO HYDROXY GROUP. R MAY BE FURTHER SUBSTITUTED IN 3 AND 5 POSITION BY CHLORINE OR BROMINE AND R&#39;&#39; IS FURTHER SUBSTITUTED IN 4&#39;&#39; OR 5&#39;&#39; POSITION BY TRIFLUOROMETHYL, ALKYL, CYCLOALKYL, ARYL AND ARALKYL RESIDUES.

asb iosl i l if 20%;)

3,646,199 SALlCYtCLlC ACID o-HYDROXYPHlENYLAMIDES FOR COMBATING BACTERIA AND FUNGI Max Schelienbaum, Muttenz, and Max Duennenberger, Frenkendorf, Switzerland, assignors to Cilia Limited, Basel, Switzerland No Drawing. Continuation-impart of application Ser. No. 745,100, July 6, 1968, This application Sept, 23, 1970, Ser. No. 74,839 Claims priority, application Switzerland, July :26, 1967, 10,607/67 Int. Cl. A0ln 9/24 US, iCl, 424-4230 6 Claims ABSTRACT ()F THE DISCLOSURE CROSS-REFERENCE This application is a continuation-in-part of applicants copending application Ser. No, 745,100, filed July 6, 1968 now US. Pat, 3,576,869, the disclosure of which is relied on. and incorporated by reference in this application,

The present invention provides a method for combating harmful microorganisms, which comprises applying to a desired site an effective amount of a salicylic acid o-hydroxyphenylamide of the formula wherein X denotes a hydrogen atom or a halogen atom and Y denotes a trifiuoromethyl group, an alkyl group having at most 12 carbon atoms, a cycloalkyl residue, a phenyl residue or an aralkyl residue, with an alkyl group Y containing at least: 8 carbon atoms if X represents hydrogen,

These salicylic acidv o-hydroyphenylamides of formula (l)-[subsequently partly also simply called salicylamides] thus correspond to one of the formulae Here X, denotes a halogen atom, preferably a chlorine or bromine atom, Y denotes a trifluoromethyl group, an alkyl group having at most 12 carbon atoms, a cycloalkyl residue, a phenyl residue or an aralkyl residue and Y denotes a trifiuoromethyl group, an alkyl group hav= Patented Feb, 3%?2 ing 8 to 12 carbon atoms, a cycloalkyl residue, a phenyl residue or an aralkyl residue The substituent Y, or Y or Y is for example in the p-position to the --HN- group and preferably in the p-position to the HO group if 1" is an alkyl group, then the latter may be unbranched or branched. n-Octyl, n-dodecyl and l,l,3,3-tetramethylbutyl groups, and in the case where the salicylic acid residue contains 2 halogen atoms, methyl, ethyl, n-propyl, isopropyl and tertiary butyl groups, may be mentioned as examples. Possible cycloalkyl residues are primarily cyclohexyl residues and possible aralkyl. residues for ex ample beuzyl residues or especially 1,1dirnethyll -phenyly methyl residues (cumyl residues),

The saiicylamides of Formula 1 may appro 'rriately be manufactured by condensation of salicylic acid halides of formula (H -Halogen especially salicylic acid chloride of the chlorides of 3,5 dibromosalicylic acid or 3,5-dichlorosalicylic acid with o-hydroxyaminobenzenes of formula wherein X and Y have the significance mentioned. it is ad vantageous to work in an inert organic solvent and with the addition of an acid-binding reagent, for example of aqueous sodium hydroxide solution.

The salicylamides of Formulae 1 to 3 can be used for combatting harmful micro-organisms and using the salicyl amides, materials for combatting harmful micro-orgam isms can be manufactured and used in the usual manher which is in itself known. A particularly valuable as pect of the new materials is the broad anti-bacterial ac tion spectrum which extends both to grampositive and also to gram-negative bacteria, At the same time the fact that the salicylamides are odourless and colourless are of paricular value from the point of, view of use technique, The present invention thus also comprises their use in con1- batting pests quite generally. It is possible to use them on a very broad basis, especially for protecting organic substrates against attack by destructive and pathogenic (also phytopathogenic) microorganisms, The new salicylamides are accordingly suitable for use both as preservatives and also as disinfectants for textiles and technical products of all kinds, in plant protection, in agriculture, in veterinary medicine and in cosmetics.

The following may be selected as examples from amongst the technical products which may be preserved with the aid of the salicylamides: textile aids or finishing agents, glues, binders, mints, colour pastes or printing pastes and similar preparations based on organicand in organic dyestuffs or pigments, also including those which contain casein or other organic compounds as admix= tures. Wall and ceiling paints, for examplethose containing an albuminous colour binder, are also protected against attack by pests by adding the new compounds, it is also possible to use them to protect timber,

Textiles which have been treated in this way also show a protection against the occurrence of perspiration odour such as is occasioned by micro-organisms.

The salicylamides can also be employed as preservatives in the cellulose and paper industry, inter alia for preventing the known slime formation, caused by microorganisms, in the equipment used for the production of paper.

Furthermore detergents and cleansing agents having an excellent anti-bacterial and/ or anti-mycotic action are obtained by combining the compounds of formulae 1 t 3 with detergents and/or surface-active substances. The salicylamides may for example be incorporated into soaps or combined with soap-free detergents and/0r surfaceactive substances or with mixtures of soaps and soap-free detergent substances; their anti-microbial activity remains 1 fully preserved in these combinations.

Cleansing agents containing a compound of Formula. i may be employed in industry and in the household, and also in the food industry, for example dairies, breweries or abattoirs. The salicylamides may also be used as a constituent of preparations which serve for cleaning and/or disinfection in hospitals and in medical practice.

The action can also be utilised in preservative and dis infectant finishes of plastics. When using plasticisers it is advantageous to add the salicylamide to the plastic with the salicylamide dissolved or dispersed in the plasticiser. it is appropriate to ensure as uniform distribution in the plastic as possible. The plastics with anti-microbial properties may be used for consumer articles of all kinds in which an activity against the most diverse germs such as for example bacteria and fungi is desired, such as for example in doormats, bathroom curtains, toilet seats, foot grids in swimming baths and wall coverings. Floor polishes and furniture polishes having a disinfectant action are obtained by incorporating the materials into wax and polishing compositions.

The salicylamides of Formula 1 may be applied in the most diverse manner to textile materials which are to be protected, for example by impregnation or spraying with solutions or suspensions which contain the compounds mentioned as the active substance. Herein the active substance content may, depending on the end use, be between 1 and 30 g. of active substance per litre of treatment liquid. In most cases textile materials of both synthetic and natural origin are adequately protected against fungal and bacterial attack by a content of 0.1 to 3% of active substance. The active substance may be employed together with other textile aids such as finishing agents, creaseproof finishes and the like.

The use forms may correspond to the usual formulations of pesticides; for example materials which contain a compound of Formula. 1 may optionally also further contain additives such as carriers, solvents, diluents, dis persing agents, wetting agents or adhesives and the like, as well as other pesticides. Finally it is also possible for several compounds of Formulae l. to 3 to be simultaneously present in such materials for combatting harmful micro-organisms.

The parts specified in the examples which follow are parts by weight and the percentages are percentages by weight unless otherwise stated.

EXAMPLE 1 22.6 parts of 3,S-dichlorosalicylic acid chloride are solved in 1.00 parts by volume of acetone. The solution.

is cooled to 5 C. and a solution of 19.1 parts of Z-amino 4-cyclohexyl-phenol in a mixture of. parts by volume acetone and 100 parts by volume oi dioxane is added at this temperature. 50 parts by volume of 2 N sodium. hydroxide solution are added dropwise to the reaction mixture at 5 to 10 C. over the course of 10 minutes and the whole is then poured into ice-water. The compound of formula Melting point Number X Y (position) 111 O.

12 r H CH CH (5) 155-156 -on,Z-- orn l CH H 13.. ll. (711 (1 H (5") -146 (3ll. CH

CIT -CH 1 l H (4') 203-204 3:4 15. e II Samcas above... (5) 18S189 16..... H CH (5) 1225-1235 17. H. C F 5') ltll -1.l2

l8.. Cl --Gll l} lJ-L-lllfi 19 Cl --CII (5) 103494 20, Ol -C(Cll'i) (5) 173-175 21. Cl Ull (-ll; (5) IDLE-1.92.5

, l ----t ?---orr, --c1n on}, on

toi Hi, :5- l'lZ. 5

l l L lly? -41! 3 2!)... Bl." C]'lg"-i iil3 to) Edit-Lilli;

etr om Oll -"CU;

a0... Br s? (5") Que-20s Melting Number X Y (position) iii 3], r Br CH (5) 164-165 32 Br -CF (5) 212-213 53 C1 CH (5) 158. 5'l50.5

---( 3GH2CH 34.. U Br CH (5) 170-171 O Hifill';

35.. (/1 -Cl-I (3) 101-101.?)

36 Cl X (4) 218-210 $7 Br Same as above (4') 235-236 38.." C1 (OH2)nCH (5) 115. 5416.5

EXAMPLE 2 Determination of the minimal inhibitory concentration (MIC) against bacteria and fungi in the dilution test The determination of the MIC (minimal inhibitory con centration) is carried out according to a test derived from standard specifications, which permits an approximation to absolute minimal inhibitory values of an active sub stance.

-A 1% strength solution and an 0.3% strength solution of the active substances in dimethylsulphoxide is intro duced into test tubes with sterile brain heart infusion broth. (bacteria) or beer wort solution (fungi) and dilu tion series with progressive 10-fold dilution are made "up with the solutions. By combination of both series the following continuous dilution series is obtained: 1000, 300, 100, 30, 10 3 ppm. and so on.

The solutions are inoculated with the bacterium Staph-- ylococcus aureus or the fungi Aspergillus niger and Rhizopus nigricans. Thereafter the material is, in the case of Staphylococcus aureus incubated for 48 hours at 37 C. (bacteriostasis) and, in the case of the fungi, for 72 hours at 30 C. (fungistasis).

The minimal inhibitory values (p.p.m.) shown in the table which follows are obtained after the times mentioned.

TABLE II Inhibitory values (p.p.m.) of-- Funglstatis Bacteriostasis Staphylococcus Aspergillus Rhizopus aure'us *lriger nigrictms Compound Number:

EXAMPLE 3 Determination of the minimal inhibitory concentration (MIC) against bacteria and fungi in the gradient plate test The compounds of Formula I, in suitable formulations (for example as solutions in dirnethylsulphoxide) of a certain concentration, are mixed with warm brain heart infusion agar (bacteria) and Mycophil agar (fungi) re spectively. The liquid mixtures are cast onto a solidified wedge-shaped base agar layer and are also allowed to solidify.

The test organisms are now applied along a line at right angles to the gradient by means of a Pasteur pipette. After an incubation for 24 hours at 37 C. (bacteria) or 72 hours at 30 C. (fungi) respectively, the length of the germs which have grown on the inoculation line is measured and expressed in ppm. of active substance.

In order to manufacture an antimicrobial cake of soap, 1.2 g. of the compound of Formula 28 are added to the following mixture: 120 g. of base soap in the form of flakes 0.12 g. of. the disodium salt of ethylene--diaminetetraacetic acid (dih'ydrate), and 0.24 g. of titanium di oxide.

The soap chips obtained by rolling are powdered by means of a rapid-running stirrer and subsequently pressed into cakes of soap.

Av 5% strength solution and a .5 strength solution in sterile tapwater are manufactured with. the antimicrobial soap. 1 ml. of these solutions is added to, in each case, 4 ml. of sterile brain heart infusion broth. By progressive 10-fold dilution in each case, two series are obtained which on combination yield the following continuous dilution series: 100, 30 10, 3, 1 ppm. of active sub" stnace.

The solutions are inoculated with a bacteria Staplzyl0- caccus aureus or Echerichia coli and incubated for 24- hours at 37 C. After this time 0.05 ml. are withdrawn from the solutions by means of a pipette and allowed to run over inclined brain heart infusion agar. Solutions (bacteriostasis) as well. as agar test tubes (bactericidal effect) are then incubated for a further 24 hours at 37 C.

The minimal inhibitory concentration and minimal lethal concentration (ppm), respectively, are now determined for the solutions and the inclined agar test tubes:

TABLE IV Action against-- Staphylococcus Escheraurcus, ichia ppm. coli Baeteriostasis [48 hours] 0.1 Bactericidal effect [24 hours].. 0. 1 100 EXAMPLE The following mixture is rolled for 20 minutes at 150" L1 on a two-roll mill: 100.00 parts of polyvinyl chloride, 4525 parts of di-Z-ethylhexylphthalate, 1.5 parts of barium/cadmium laurate, 0.25 part of stearic acid and 7.75 parts of a. solution of 1.55 parts of the compound of Formula 37 in 6.25 parts of di-Z-ethylhexyl phthalate. The roll nip is so adjusted that 1 mm. thick hides are produced which are subsequently pressed for 80 minutes at 165 to 170 C. at 1400 kg./cm.

In order [0 test the action against bacteria, mm. diameter discs are punched out of the plasticized polyvinyl chloride and laid on brain heart infustion agar plates which have been pre-inoculated with Staphylococcus aureus. The plates are thereafter incubated for 24 hours at 37 The items assessed are, on the one hand, the inhibitory zone which manifests itself around the discs (Hz. in mm.) and, on the other hand, the microscopically determinable growth (W in percent) underneath and on the plasticised polyvinyl chloride:

TABLE V Soaked t W in. percent 3 Soaking: 24 hours at Cu EXAMPLE 6 Samples of 100 cotton cretonne are impregnated with a 1% strength solution of Compound No, 21 in isopropanol at C. on a padding machine and subsequently squeezed out to leave 100% liquor uptake.

Samples of 100 g. wool Cheviot are also treated in the same manner.

The fabrics dried at to 40 C, contain 1% of active substance relative to their own weight.

In order to test. the action against bacteria, 10 mm. diameter discs of the impregnated fabrics are, ansoaked and after soaking, laid for 24 hours at 29 C: onto brain. heart. infusion agar plates which are pro-inoculated with staphylococcus aureus. The plates are thereafter incubated for 24 hours atv 37 C.

The items assessed are, on the one hand, the inhibitory zone which arises around the discs (Hz. in mm.) and, on the other hand, the microscopically determinable growth W in percent) under and on the fabric:

Soaked W in percent Without soaking HZ Win 1 111111. pUl'CGlllS Substrate {with 1% active Substance) What is claimed is: it. A, method for combating harmful bacteria and fungi which. comprises applying to a desired site an effective L bacterial and fungicidal amount of a salicylic acid o-hy droxyphenylamide of the formula r 7 0 at 0 g "the method according to claim 1, wherein the saii. cylic acid o-hydroxyphenylamide has the formula (H -CH wherein. X denotes a member selected from the group consisting of chlorine and bromine.

4. The method according to claim it, wherein the salicylio acid o-hydroxyphenylamide has the formula wherein. X denotes a member selected from the group consisting of chlorine and bromine.

5. The method according to claim 1, wherein the salicylic acid o-hydroxyphenylamide has the formula X on. no

/ c NH Q as as ,x 0

1 1: cs (f cs on cs wherein X denotes a member selected from the group consisting of chlorine or bromine,

or The method according to claim Jl, wherein the sail cylio acid otiydroxypheriylamide has the formula wherein X denotes a member selected from the group consisting of chlorine and bromine.

References Cited UNITED STATES PATENTS Thompson Wow. IZS2---403 JEROME D. GOLDBERG, Primary Examiner US. C1. 

